Imperatorin Targets MCL-1 to Sensitize CD133+ Lung Cancer Cells to γδ-T Cell-Mediated Cytotoxicity.

BACKGROUND/AIMS: CD133+ cancer cells display low sensitivity to anti-cancer treatment; thus, combination treatment with adjuvant drugs is required to improve the efficiency of cancer therapy. The aim of this study was to explore the effect of imperatorin, a linear furanocoumarin compound, on γδ T cell-mediated cytotoxicity against CD133+ lung cancer cells. METHODS: CD133+ and CD133- subgroups from A549 and PC9 lung cancer cells were sorted by using flow cytometry. The cytotoxicity of γδ T cells against cancer cells was evaluated by measuring lactate dehydrogenase release. The concentration of tumor necrosis factor-related apoptosis-inducing ligand in the co-culture system was determined by using an enzyme-linked immunosorbent assay. Mitochondrial membrane potential, expression of death receptor 4 (DR4) and DR5 on the cell surface, and rate of apoptosis were measured by flow cytometry. Cytochrome c release and cellular protein expression were detected by western blot analysis. RESULTS: Compared with CD133- cells, CD133+ cells were resistant to γδ T cell-mediated cytotoxicity. However, imperatorin significantly increased the sensitivity of CD133+ lung cancer cells to γδ T cell treatment in vitro and in vivo. Mechanically, we found that myeloid cell leukemia 1 (MCL-1), an important anti-apoptotic protein belonging to the Bcl-2 family, was overexpressed in CD133+ A549 and PC9 cells compared to their corresponding CD133- cells. Co-treatment with imperatorin and γδ T cells suppressed the expression of MCL-1, and thus promoted the mitochondrial apoptosis mediated by γδ T cells in CD133+ A549 and PC9 lung cancer cells. CONCLUSION: Up-regulated MCL-1 in CD133+ lung cancer cells is responsible for their resistance to γδ T cells. Furthermore, the combination of γδ T cells with imperatorin sensitized CD133+ lung cancer cells to γδ T cell-mediated cytotoxicity by targeting MCL-1.

Comparison of the upper and lower airway microbiome in early postoperative lung transplant recipients.

The upper and lower respiratory tract may share microbiome because they are directly continuous, and the nasal microbiome contributes partially to the composition of the lung microbiome. But little is known about the upper and lower airway microbiome of early postoperative lung transplant recipients (LTRs). Using 16S rRNA gene sequencing, we compared paired nasal swab (NS) and bronchoalveolar lavage fluid (BALF) microbiome from 17 early postoperative LTRs. The microbiome between the two compartments were significantly different in Shannon diversity and beta diversity. Four and eight core NS-associated and BALF-associated microbiome were identified, respectively. NS samples harbored more Corynebacterium, Acinetobacter, and Pseudomonas, while BALF contained more Ralstonia, Stenotrophomonas, Enterococcus, and Pedobacter. The within-subject dissimilarity was higher than the between-subject dissimilarity, indicating a greater impact of sampling sites than sampling individuals on microbial difference. There were both difference and homogeneity between NS and BALF microbiome in early postoperative LTRs. High levels of pathogens were detected in both samples, suggesting that both of them can reflect the diseases characteristics of transplanted lung. The differences between upper and lower airway microbiome mainly come from sampling sites instead of sampling individuals. IMPORTANCE: Lung transplantation is the only therapeutic option for patients with end-stage lung disease, but its outcome is much worse than other solid organ transplants. Little is known about the NS and BALF microbiome of early postoperative LTRs. Here, we compared paired samples of the nasal and lung microbiome from 17 early postoperative LTRs and showed both difference and homogeneity between the two samples. Most of the "core" microbiome in both NS and BALF samples were recognized respiratory pathogens, suggesting that both samples can reflect the diseases characteristics of transplanted lung. We also found that the differences between upper and lower airway microbiome in early postoperative LTRs mainly come from sampling sites instead of sampling individuals.

Erlotinib versus chemotherapy as first-line treatment for patients with advanced EGFR mutation-positive non-small-cell lung cancer (OPTIMAL, CTONG-0802): a multicentre, open-label, randomised, phase 3 study.

BACKGROUND: Activating mutations in EGFR are important markers of response to tyrosine kinase inhibitor (TKI) therapy in non-small-cell lung cancer (NSCLC). The OPTIMAL study compared efficacy and tolerability of the TKI erlotinib versus standard chemotherapy in the first-line treatment of patients with advanced EGFR mutation-positive NSCLC. METHODS: We undertook an open-label, randomised, phase 3 trial at 22 centres in China. Patients older than 18 years with histologically confirmed stage IIIB or IV NSCLC and a confirmed activating mutation of EGFR (exon 19 deletion or exon 21 L858R point mutation) received either oral erlotinib (150 mg/day) until disease progression or unacceptable toxic effects, or up to four cycles of gemcitabine plus carboplatin. Patients were randomly assigned (1:1) with a minimisation procedure and were stratified according to EGFR mutation type, histological subtype (adenocarcinoma vs non-adenocarcinoma), and smoking status. The primary outcome was progression-free survival, analysed in patients with confirmed disease who received at least one dose of study treatment. The trial is registered at ClinicalTrials.gov, number NCT00874419, and has completed enrolment; patients are still in follow-up. FINDINGS: 83 patients were randomly assigned to receive erlotinib and 82 to receive gemcitabine plus carboplatin; 82 in the erlotinib group and 72 in the chemotherapy group were included in analysis of the primary endpoint. Median progression-free survival was significantly longer in erlotinib-treated patients than in those on chemotherapy (13.1 [95% CI 10.58-16.53] vs 4.6 [4.21-5.42] months; hazard ratio 0.16, 95% CI 0.10-0.26; p<0.0001). Chemotherapy was associated with more grade 3 or 4 toxic effects than was erlotinib (including neutropenia in 30 [42%] of 72 patients and thrombocytopenia in 29 [40%] patients on chemotherapy vs no patients with either event on erlotinib); the most common grade 3 or 4 toxic effects with erlotinib were increased alanine aminotransferase concentrations (three [4%] of 83 patients) and skin rash (two [2%] patients). Chemotherapy was also associated with increased treatment-related serious adverse events (ten [14%] of 72 patients [decreased platelet count, n=8; decreased neutrophil count, n=1; hepatic dysfunction, n=1] vs two [2%] of 83 patients [both hepatic dysfunction]). INTERPRETATION: Compared with standard chemotherapy, erlotinib conferred a significant progression-free survival benefit in patients with advanced EGFR mutation-positive NSCLC and was associated with more favourable tolerability. These findings suggest that erlotinib is important for first-line treatment of patients with advanced EGFR mutation-positive NSCLC. FUNDING: F Hoffmann-La Roche Ltd (China); Science and Technology Commission of Shanghai Municipality.

Homologous recombination repair gene mutations as a predictive biomarker for immunotherapy in patients with advanced melanoma.

Background: Nowadays, immunotherapy targeting immune checkpoint receptors is one of the cornerstones of systemic treatment in melanoma. Homologous recombination repair (HRR) is one of the DNA damage response (DDR) pathways, which has been proved to correlate with the efficacy of platinum-based chemotherapy, PARP inhibitor therapy, and immunotherapy in a variety of cancers. However, their predictive value of HRR remained unknown in patients with advanced melanoma. Methods: Data of advanced melanoma patients from an independent cohort (Samstein2018) were used to analyze the correlation with immunogenic markers and the prognostic effect of HRR on immunotherapy, and another four cohorts (pooled cohort: Miao2018, Allen 2015, Hugo2016, and Synder2014) were used for validation. Immune infiltration cell scores analyzed by TCGA-SKCM cohort were used to explore potential mechanisms related to the immune microenvironment. Results: Compared to patients with an HRR wild type (HRRwt), those with HRR mutations (HRRmut) in anti-CTLA-4-treated patients of the Samstein2018 cohort had higher tumor mutation burden (TMB; P = 0.0041) and longer median overall survival (mOS; P = 0.0094). In terms of results validation, it was also confirmed that the mOS (P = 0.0014) of HRRmut patients receiving anti-CTLA-4 therapy was significantly better than that of HRRwt patients in the pooled cohort, and objective response rates (ORR; P = 0.0053) were also found to be significant. However, there was no significant difference in mOS between HRRmut patients who received anti-PD-1/L1 therapy and HRRwt patients in either the discovery (Samstein2018 cohort, P = 0.94) or validation (pooled cohort, P = 0.96) set. Exploratory analysis found that although HRRmut patients showed no significant difference in mOS between anti-CTLA-4 and anti-PD-1/L1 therapy (P = 0.79), the mOS value of the anti-CTLA-4 therapy group (31.7 months) in HRRmut patients was numerically superior to the anti-PD-1/L1 therapy group (27.5 months). In contrast, the mOS of the anti-CTLA-4 therapy group was significantly lower than that of the anti-PD-1/L1 therapy group (12.4 vs. 32.0 months) in HRRwt patients. In addition, transcriptome profiling analysis revealed that the 29 (65.9%)-gene mutation of the HRR pathway associated with reshaping of the immunological microenvironment in melanoma. Conclusions: HRR mutations were associated with a higher TMB level, and better anti-CTLA-4 therapy outcomes. HRR may serve as an independent predictor of anti-CTLA-4 therapy efficacy in patients with advanced melanoma and their clinical value warrants further investigation.

Sleep quality, anxiety and depression in advanced lung cancer: patients and caregivers.

OBJECTIVE: To investigate the clinical implications of sleep quality, anxiety and depression in patients with advanced lung cancer (LC) and their family caregivers (FCs). METHODS: A total of 98 patients with advanced LC and their FCs (n=98) were recruited from the Oncology Department in Nanfang Hospital. The Pittsburgh Sleep Quality Index (PSQI), consisting of seven components that evaluate subjective sleep quality, sleep latency, duration of sleep, sleep efficiency, sleep disturbances, sleep medication usage and daytime dysfunction, was used to assess sleep quality. Using the tool of Zung Self-rating Anxiety Scale (SAS) and Zung Self-rating Depression Scale (SDS), we tested the patients' status of anxiety and depression, respectively. RESULTS: The prevalences of poor sleep quality, anxiety and depression in patients were 56.1%, 48.9% and 56.1%, respectively, while those in FCs were 16.3%, 32.6% and 25.5%, respectively. Patients had higher PSQI, SAS and SDS scores than did FCs (p<0.05). Significant correlations were found between the patients' and FCs' scores of PSQI/SAS/SDS (p<0.05). Multivariate Cox regression analyses indicated that sleep disturbances in patients (HR 0.413, 95% CI 0.21 to 0.80, p=0.01) and the global PSQI score of FCs (HR 0.31, 95% CI 0.14 to 0.71, p=0.00) were independent risk factors for patients' first-line progression-free survival (PFS). Moreover, patients' sleep latency (HR 2.329, 95% CI 1.36 to 3.96, p=0.00) and epidermal growth factor receptor mutations (HR 1.953, 95% CI 1.12 to 3.38, p=0.01) were significant prognostic factors for their overall survival (OS). CONCLUSIONS: We demonstrated that presence of sleep disturbances in patients with advanced LC and the global PSQI Score of their FCs may be risk predictors for patients' poor first-line PFS. Patients' sleep latency was a potential risk factor for their OS.

The Airway Microbiota Signatures of Infection and Rejection in Lung Transplant Recipients.

Infection and rejection are the two most common complications after lung transplantation (LT) and are associated with increased morbidity and mortality. We aimed to examine the association between the airway microbiota and infection and rejection in lung transplant recipients (LTRs). Here, we collected 181 sputum samples (event-free, n = 47; infection, n = 103; rejection, n = 31) from 59 LTRs, and performed 16S rRNA gene sequencing to analyze the airway microbiota. A significantly different airway microbiota was observed among event-free, infection and rejection recipients, including microbial diversity and community composition. Nineteen differential taxa were identified by linear discriminant analysis (LDA) effect size (LEfSe), with 6 bacterial genera, Actinomyces, Rothia, Abiotrophia, Neisseria, Prevotella, and Leptotrichia enriched in LTRs with rejection. Random forest analyses indicated that the combination of the 6 genera and procalcitonin (PCT) and T-lymphocyte levels showed area under the curve (AUC) values of 0.898, 0.919 and 0.895 to differentiate between event-free and infection recipients, event-free and rejection recipients, and infection and rejection recipients, respectively. In conclusion, our study compared the airway microbiota between LTRs with infection and acute rejection. The airway microbiota, especially combined with PCT and T-lymphocyte levels, showed satisfactory predictive efficiency in discriminating among clinically stable recipients and those with infection and acute rejection, suggesting that the airway microbiota can be a potential indicator to differentiate between infection and acute rejection after LT. IMPORTANCE Survival after LT is limited compared with other solid organ transplantations mainly due to infection- and rejection-related complications. Differentiating infection from rejection is one of the most important challenges to face after LT. Recently, the airway microbiota has been reported to be associated with either infection or rejection of LTRs. However, fewer studies have investigated the relationship between airway microbiota together with infection and rejection of LTRs. Here, we conducted an airway microbial study of LTRs and analyzed the airway microbiota together with infection, acute rejection, and clinically stable recipients. We found different airway microbiota between infection and acute rejection and identify several genera associated with each outcome and constructed a model that incorporates airway microbiota and clinical parameters to predict outcome. This study highlighted that the airway microbiota was a potential indicator to differentiate between infection and acute rejection after LT.

Comparison of AAV/IL-7 autocrine (T cell) versus paracrine (DC) gene delivery for enhancing CTL stimulation and function.

Adoptive transfer of antigen-specific cytotoxic T lymphocyte (CTL) into patients holds promise in treating cancer. Such anti-cancer CTL are stimulated by professional antigen-presenting dendritic cells (DC). We hypothesize the gene delivery of various Th1-response cytokines, such as interleukin 7 (IL-7), should further enhance CTL stimulation and activity. However, the issue as to which cell type, DC (paracrine) or the T cell (autocrine), should express a particular Th1 cytokine gene for optimal CTL stimulation has never been addressed. We used adeno-associated virus-2 (AAV) to compare delivery of IL-7 and IL-2 genes into DC or T cells and to exogenous commercial cytokines for generating robust carcinoembryonic antigen (CEA)-specific CTL. AAV/IL-7 transduction of T cells (autocrine delivery) generated CTL with the highest killing capability. Consistent with this, AAV/IL-7 delivery generated T cell populations with the highest proliferation, highest interferon gamma expression, highest CD8(+):CD4(+) ratio, highest CD8(+), CD69(+) levels, and lowest CD4(+), CD25(+) (Treg) levels. These data are consistent with higher killing by the AAV/IL-7-altered CTL. These data strongly suggest that IL-7 autocrine gene delivery is optimal for CTL generation. These data also suggest Th1 cytokine autocrine versus paracrine delivery is an important issue for immuno-gene therapy and uncovers new questions into cytokine mechanism of action.

Expression of miR-21 and miR-138 in colon cancer and its effect on cell proliferation and prognosis.

Effect of miR-21 and miR-138 on the proliferation of colon cancer cells and their association with prognosis were investigated. Expression levels of miR-21 and miR-138 in colorectal cancer and normal adjacent tissues were compared. Differential expression of miR-21 and miR-138 in colon cancer tissues with different clinicopathological features were analyzed. miR-21 and miR-138 expression vectors were established and transfected into cells of colon cancer cell line SW480. Methyl thiazolyl tetrazolium (MTT) assay was used to detect the proliferation of SW480 cells. Kaplan-Meier method and log-rank test were used to study the relationship between miR-21 and miR-138 expression and prognosis. Cox proportional hazards model was used to analyze the factors related to prognosis of colon cancer. Expression level of miR-21 in colon cancer tissues was significantly higher than that in adjacent tissues, and expression level of miR-138 was lower in cancer tissues than in adjacent tissues (P<0.001). Expression of miR-21 and miR-138 was associated with the degree of differentiation, lymph node metastasis, distant metastasis, and TNM stage (P<0.05). miR-21 promotes proliferation of colon cancer cell line SW480, and miR-138 inhibits cell proliferation. Survival analysis showed that the survival time of patients with high expression of miR-21 was significantly shorter than that of patients with low expression of miR-21, while survival time of patients with high expression of miR-138 was significantly longer than that of patients with low expression of miR-138 (log-rank, P<0.05). miR-21 is highly expressed in colon cancer tissues and is positively associated with the degree of malignancy of patients but negatively associated with survival. miR-138 expression is low in colon cancer tissues and is negatively associated with the degree of malignancy of patients but positively associated with survival. miR-21 and miR-138 may be involved in the regulation of colon cancer cell proliferation.

Clinical applications of liquid biopsies for early lung cancer detection.

Over the past decade, the clinical utility of liquid biopsies in lung cancer has drawn increasing attention. Having been successfully applied in targeted therapy for late stage lung cancer, liquid biopsies are being further investigated regarding their potential role for early detection of lung cancer. Novel biomarkers with high sensitivity and specificity are crucial for identifying patients at early stages as well as for monitoring high-risk populations. A variety of bodily fluids (such as plasma, serum, and sputum) and biomarkers (such as cfDNA, CTCs, gene methylation, and miRNA) have been investigated for their potential role in the diagnosis of lung cancer. In this review, we summarize recent advances in circulating biomarkers regarding the early detection of lung cancer and discuss their potential applications and challenges in clinical settings.

Prognostic Implications of Tripartite Motif Containing 24 Expression Levels in Patients with Solid Tumors: A Systematic Review and Meta-Analysis.

Objective: To systematically investigate the prognostic implications of tripartite motif containing 24 (Trim 24) expression levels in Patients with solid tumors. Materials and Methods: Pubmed, Embase, China National Knowledge Infrastructure, and Wanfang databases were searched through December 2017 to identify studies examining the relationship between Trim 24 expression levels and outcomes in solid tumor patients. The hazard ratios (HRs) with corresponding 95% confidence intervals were used to evaluate the association between Trim 24 and overall survival (OS). Results: Ten studies with 1370 patients were included. The overall pooled prevalence for Trim 24 overexpression was 59.0% (p < 0.01). Moreover, the pooled HR of Trim 24 for OS was 0.43 (p = 0.04) by univariate analysis in 10 articles (1370) and 0.62 (p = 0.08) by multivariate analysis in 5 studies (845). Trim 24 over-expression was associated with tumor invasiveness (odds ratio [OR] = 2.05, p < 0.01) and tumor-node-metastasis stage (OR = 2.42, p = 0.03). Conclusions: This study demonstrated that Trim 24 expression levels may be a useful prognostic biomarker in patients with solid tumors.

Use and specificity of breast cancer antigen/milk protein BA46 for generating anti-self-cytotoxic T lymphocytes by recombinant adeno-associated virus-based gene loading of dendritic cells.

Antigen-targeted immunotherapy is an emerging treatment for breast cancer. However, useful breast cancer antigens are only found in a subset of cancer patients. BA46, also known as lactadherin, is a membrane-associated glycoprotein that is expressed in most breast cancer cells but not in general hematopoietic cell populations. Moreover, it is much more difficult to generate CTLs against self-antigens. We wished to determine if the use of recombinant adeno-associated virus (rAAV) type 2 vectors for gene-loading of dendritic cells (DCs) could generate rapid, effective cytotoxic T lymphocytes (CTLs) against BA46. We were able to demonstrate that AAV/BA46/Neo-loading of DCs resulted in: (1) BA46 expression in DCs, (2) chromosomal integration of the AAV/BA46/Neo vector within DCs, (3) strong, rapid BA46-specific, MHC class I-restricted CTLs in only 1 week, (4) T-cell populations with significant interferon-gamma (IFN-gamma) expression but low IL-4 expression, (5) high CD80 and CD86 expression in DCs, and (6) high CD8:CD4 and CD8:CD56 T cell ratios. These data suggest that rAAV-loading of DCs may be useful for immunotherapeutic protocols against self-antigens in addition to viral antigens and that the BA46 antigen is potentially appropriate for cell-mediated immunotherapeutic protocols addressing ductal breast cancer.

[Immunostimulatory effect of human peripheral blood dendritic cells transfected by adeno-associated virus containing Her2/neu gene].

OBJECTIVE: To observe the immunostimulatory effect of human peripheral blood dendritic cells (DCs) transfected by Her2/neu gene delivered by adeno-associated virus. METHODS: The mononuclear cells in healthy donors were isolated by Ficol-Hypaque density gradient separation and divided into transfection group and control group without transfection by the recombinant virus. The cells were initially cultured in RPMI 1640 medium supplemented with 10% AB human serum, followed by addition of granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4 and tumor necrosis factor (TNF)-alpha into the medium. The surface markers of DC were detected by flow cytometry, allogeneic mixed lymphocyte reaction assayed by 3H-TdR incorporation and the specific killing activity of T cells evaluated by MTT assay. RESULTS: The expression rates of CD1a, CD86 and CD83 of the transfected DCs were 98.10%, 99.42%, and 84.59%, and those of the non-transfected DCs were 92.69%, 98.07%, and 82.72%, respectively, showing no obvious differences between the two groups. The rates of CD4 and CD80 expression were 61.02% and 97.61%, respectively, in the transfected DCs, significantly higher than those in the non-transfected cells (36.19% and 55.5%). Both groups of DCs stimulated a strong T cell proferation response. The transfected DCs were capable of inducing specific killing of the target tumor cells, with the highest killing rate of (39.7+/-7.2)%. CONCLUSION: The immunostimulatory effect of human peripheral blood DCs can be enhanced by Her2/neu gene transfection mediated by adeno-associated virus.

[Cytotoxicity of cytotoxic T lymphocytes induced by keratin 19-sensitized dendritic cells against MCF-7 cells in vitro].

OBJECTIVE: To investigate the cytotoxicity of cytotoxic T lymphcytes (CTL) induced by keratin 19 (K19)-sensitized dendritic cells (DC) against MCF-7 cells in vitro. METHODS: DC isolated from peripheral blood mononuclear cells were cultured in vitro and sensitized by K19 peptide to generate specific CTL. The phenotypes and intracytoplasm cytokine of DC were analyzed by flow cytometry. Mixed leukocyte responses were evaluated by (3)H-TdR incorporation assay. The T cells were generated by DC pulsed with K19 antigen and T cell cytotoxicity was measured by (51)Cr release assay. RESULTS: The DCs derived from peripheral blood mononuclear cells expressed high levels of CD40, CD86, CD80 and CD83. Mixed leukocyte responses induced by the DCs pulsed with K19 was stronger than that induced by naive DCs (P<0.01). The cytotoxicity rate of CTL induced by the sensitized DCs was higher than that of CTL induced by naive DCs (P<0.01). The cytotoxity activity was enhanced by increasing the effector/target ratio (P<0.01). CONCLUSION: After sensitization with K19 antigen, the DCs can stimulate T cell proliferation and induce cytotoxicity activity against MCF-7 cells. Increased effector/target ratio may enhance the cytotoxity activity. Sensitized DCs possess the potential for amplification in immunotherapy of carcinoma.

Role of Aurora-A in Ovarian Cancer: A Meta-Analysis.

BACKGROUND: Recently, several studies have examined associations between Aurora-A expression and clinical outcome in patients with ovarian cancer, but yielded conflicting results with respect to survival. Therefore, the aim of this study was to evaluate the prognostic significance of Aurora-A in ovarian cancer by performing a meta-analysis. METHODS: PubMed, Cochrane library, Web of Science, Embase, Medline and Chinese BioMed Database (CBM) databases were searched systematically and only articles in which Aurora-A expression was detected by immunohistochemical staining were included. Hazard ratios (HRs) with 95% confidence intervals (CIs) were extracted and pooled for overall survival (OS) and disease-free survival (DFS). RESULTS: Our results show that the pooled HR for OS was 1.40 (95% CI 0.82-1.98, p < 0.01) by univariate analysis in 7 articles (1,028) and 0.32 (95% CI 0.04-0.615, p = 0.23) by multivariate analysis in 3 studies (155). The association between Aurora-A expression and DFS was also statistically significant in 5 studies (HR = 1.14, 95% CI 0.50-1.78, p < 0.01). CONCLUSION: This present meta-analysis suggests that the Aurora-A expression may be associated with poor prognosis in patients with ovarian cancer. Furthermore, studies of larger scale and well-matched regimes are warranted to confirm the findings in the future.

CCNB2 overexpression is a poor prognostic biomarker in Chinese NSCLC patients.

Cyclin B2 (CCNB2), a member of cyclin family proteins, serves a key role in progression of G2/M transition. The clinical value of CCNB2 in non-small cell lung cancer is still unknown. The aim of our study is to identify the role of CCNB2 in NSCLC patients. The status of CCNB2 in NSCLC tissues and normal lung tissues was observed in Gene Expression Omnibus (GEO: GSE19804). CCNB2 mRNA and protein expressions were detected in NSCLC and normal lung tissues by using Real-time PCR and immunohistochemistry. The association of CCNB protein expression with clinical characteristics of 186 NSCLC patients was analyzed by immunohistochemistry. Based on microarray data (GEO: GSE19804), we observed that CCNB2 was significantly overexpressed in NSCLC tissues compared with paired adjacent normal lung tissue. Furthermore, we verified mRNA and protein levels of CCNB2 expression were both increased in NSCLC tissues. We found high levels of CCNB2 protein were positively associated with the status of differentiated degree, tumor size, lymph node metastasis, distant metastasis, and clinical stage. Meanwhile, CCNB2 protein overexpression was an independent unfavorable prognostic factor for the overall survival of patients with NSCLC. In conclusion, overexpression of CCNB2 protein is associated with clinical progression and poor prognosis in NSCLC.

[Clinical observation of the efficacy of navelbine combined with ifosfamide and cisplatin in the treatment of advanced breast cancer].

OBJECTIVE: To observe the efficacy of navelbine combined with ifosfamide and cisplatin in the treatment of advanced breast cancer. METHODS: Forty cases of pathologically confirmed stages III-IV breast cancer were treated with combined chemotherapy including intravenously infused navelbine (25 mg/m2, days 1 and 8), ifosfamide (1.3 g/m2, days 1 to 5) and cisplatin (20 mg/m2, days 1-5), administered in cycles of 21 days. The therapeutic effect was assessed after at least two cycles were completed. RESULTS: Complete remission was achieved in 6 cases and partial remission in 16, with an overall responding rate of 55%. The main toxicity was bone marrow suppression manifested by grades III-IV leukopenia occurring at a rate of 40%. CONCLUSION: Navelbine in combination with ifosfamide and cisplatin is effective and safe in the treatment of advanced breast cancer.

Emergency management of respiratory failure by endotracheal intubation through the nose under the guidance of portable bedside fibrobronchoscope.

OBJECTIVE: To investigate the value of portable fibrobronchoscope in the management of respiratory failure by endotracheal intubation through the nose. METHODS: Fifty-eight patients with acute or chronic respiratory failure received mechanical ventilation by endotracheal intubation through the nose under the guidance of portable fibrobronchoscope. RESULTS: Intubation was successfully performed in all the patients in a single attempt (which took 30 min to 5 min) without interruption of autonomous breathing or incurring laryngospasm or cardiac arrest. After mechanical ventilation for 30 min, conspicuous improvement of respiratory failure was observed in all the cases. CONCLUSION: With convenient and easy manipulation, portable fibrobronchoscope provides quick and accurate guidance for endotracheal intubation through the nose in the emergency management of respiratory failure.

Preparation and identification of monoclonal antibodies against human brain-derived neurotrophic factor.

OBJECTIVE: To prepare monoclonal antibodies (mAbs) against human brain-derived neurotrophic factor (BDNF). METHODS: BALB/C mice were immunized with purified BDNF protein in conjunction with acid-treated Salmonella (antigen:thallus=1:5), and mAbs were subsequently derived by hybridoma technique. RESULTS: Three hybridoma cell lines secreting anti-BDNF mAbs were obtained, designated as B1, B2 and 4D1 respectively and all categorized into IgG1 subtype. The titers of the mAbs in the ascitic fluid of the rats ranged from 1x10(6) to 1x10(5), and their relative affinities were B2>B1>4D1. CONCLUSION: mAbs against BDNF are successfully prepared, which can be instrumental for further study of the expression and distribution of BDNF in vivo, and the detection of BDNF produced by genetic engineering.

[Clinical study of serum tissue polypeptide-specific antigen in breast cancer].

OBJECTIVE: To investigate the expression of serum tissue polypeptide-specific antigen (TPS) in breast cancer patients and its clinical value in such cases. METHODS: Altogether 160 subjects (90 patients with breast cancer, 40 with benign breast lesions, and 30 healthy subjects) were enrolled in this study. The serum TPS and CA153 levels were measured by ELISA in all the subjects. RESULTS: The levels and positivity rate of serum TPS and CA153 in breast cancer group were significantly higher than those in the normal subjects group and benign lesion group (P<0.01), and became even higher as the malignancy progressed. High serum TPS level was detected in the cancer patients in stage I. Serum TPS level was the most sensitive to bone metastasis of the malignancy, but its highest levels occurred in cases of lymphoid node metastasis (P<0.05). In patients who responded favorably to the treatment, serum TPS and CA153 levels were significantly reduced (P<0.05), but the reduction in TPS levels tended to be more obvious (P<0.05). CONCLUSION: Serum TPS can be used as a very useful and sensitive tumor marker in the diagnosis of breast cancer, especially in case of bone metastasis, and may be of great value in clinical decision-making and assessment of therapeutic effect.

[89SrCl2 and/or Bonefos in the treatment of bone metastasis from pulmonary carcinoma].

BACKGROUND: To evaluate the effect of 89SrCl2 and/or Bonefos in the treatment of bone metastasis from pulmonary carcinoma. METHODS: A total of sixty-seven lung cancer patients with bone metastasis were enrolled in this study, who were divided into three groups: nineteen cases were treated with 89SrCl2; twenty-eight cases with Bonefos; and twenty cases combined 89SrCl2 with Bonefos. RESULTS: The total relief rate of the bone pain: the 89SrCl2 group was 84.2%, and the Bonefos group was 80.4%, the combination group was 90.0%. There was no statistical difference among three groups (P > 0.05). The effective rate of the bone metastasis: the 89SrCl2 group was 15.7%, the Bonefos group was 10.7%, and the combination group was 45.0%. The combination group had significantly higher effective rate than that of the 89SrCl2 group or the Bonefos group alone (P < 0.05). The rate of improvement of quality of life: the 89SrCl2 group was 47.3%, the Bonefos group was 42.8%, and the combination group was 80.0%. The combination group had significantly higher effective rate than that of the 89SrCl2 group or the Bonefos group alone (P < 0.05). The side effects of three groups were minimal. CONCLUSIONS: 89SrCl2 and Bonefos are two effective and safe drugs on relief of pain. Combined 89SrCl2 and Bonefos might be a better therapy for bone metastasis and improvement of quality of life than the single one, and the side effect is slight and tolerable.